Dasatinib Promotes Chondrogenic Differentiation of Human Mesenchymal Stem Cells via the Src/Hippo-YAP Signaling Pathway

Abstract

Mesenchymal stem cells (MSCs) are progenitors of chondrocytes and could be used as a potential therapy for cartilage defects in diarthrodial joints. However, promoting chondrogenic differentiation of MSCs remains a daunting challenge. As a small molecular drug, dasatinib can promote MSC differentiation, although the exact mechanisms of chondrogenic differentiation are unclear. In this study, the differentiation of MSCs into osteoblasts, adipocytes, and chondrocytes was assessed by the protein and mRNA levels of osteoblast- and chondrocyte-related proteins using western blotting and real-time polymerase chain reaction, respectively. MSCs were induced to differentiate into chondrocytes or osteoblasts with or without dasatinib in vitro. The effects of dasatinib on cartilage regeneration were also assessed in vivo in a rabbit model of full-thickness cartilage defects using methacrylate gelatin hydrogel as scaffolds. Dasatinib promoted chondrogenic differentiation and inhibited osteogenic differentiation of MSCs. Furthermore, dasatinib significantly inhibited the expression of YAP and TAZ and the phosphorylation of Src, but it enhanced serine phosphorylation of YAP during the chondrogenic differentiation of MSCs in vitro. Inhibition of the Hippo pathway using XMU-MP-1 dramatically suppressed the serine phosphorylation of YAP and chondrogenic differentiation of MSCs. Moreover, we confirmed that the sustained release of dasatinib from the hydrogel promoted rabbit cartilage repair. The results demonstrated that dasatinib might promote chondrogenic differentiation of MSCs via the Src/Hippo-YAP signaling pathway and that hydrogel sustained-release dasatinib had a certain effect on the repair of cartilage defects.

Yes-Associated Protein 1: Role and Treatment Prospects in Orthopedic Degenerative Diseases

“…Several studies have confirmed that YAP1 is overexpressed in OA animal model tissues, and the YAP1 content increases with the severity of OA (Nie et al, 2019;Zhang et al, 2019;Kania et al, 2020). Gong et al (2019) reached the same conclusion in studies with an OA mouse model and human OA tissue:…”

“…Several studies have confirmed that YAP1 is overexpressed in OA animal model tissues, and the YAP1 content increases with the severity of OA ( Nie et al, 2019 ; Zhang et al, 2019 ; Kania et al, 2020 ). Gong et al (2019) reached the same conclusion in studies with an OA mouse model and human OA tissue: (1) YAP1 transcription was upregulated in knee chondrocytes of OA mice by RT-qPCR analysis; (2) YAP1 levels detected in articular chondrocytes of OA mice were upregulated by Western blot analyses; (3) and YAP1 mRNA and protein levels were increased in human chondrocytes of OA compared with normal human cartilage.…”

“…YAP1 negatively regulates chondrocyte differentiation by activating the β-catenin signaling pathway, and YAP1 overexpression enhances chondrocyte proliferation but inhibits chondrocyte differentiation ( Yang et al, 2017 ). In another study of cartilage differentiation ( Nie et al, 2019 ), dasatinib treatment was observed to significantly upregulate the serine phosphorylation of YAP1 and downregulate the protein expression of YAP1 and TAZ in a dose-dependent manner, promoting cartilage gene differentiation of MSCs by promoting the serine phosphorylation of YAP1. Goto et al (2018) also showed that YAP1/TAZ activation in chondrogenesis can inhibit chondrocyte differentiation and maturation, leading to chondrodysplasia through a mechanism in which the activation of YAP1 or TAZ leads to decreased Sox9 expression.…”

“…Mouse MSCs The increased expression of YAP1 in the nucleus promotes the osteogenesis of MSCs, inhibits their adipogenic differentiation, and leads to dedifferentiation of chondrocytes Bian et al, 2016 Mouse osteoblast The regulation of OB by YAP1/TAZ is closely associated with the developmental stage of individuals and the differentiation state of cells Xiong et al, 2018 Mouse osteoblast Piezo1 in osteoblasts promotes the expression of COL2α1 and COL9α2 in osteoblast cell lines through YAP1 Wang et al, 2020 Mouse osteoclast YAP1 is an essential regulator for osteoclast differentiation and activity Zhao et al, 2018 the intra-articular injection of verteporfin, a selective inhibitor of YAP1, can significantly maintain cartilage homeostasis in a mouse OA model and that a sustained-release system consisting of chitosan microspheres and verteporfin has significant anti-OA effects (Zhang et al, 2020). Another YAP1 activity inhibitor, dasatinib, can also perform a similar function (Nie et al, 2019). Dasatinib can significantly inhibit YAP1 expression, promote cartilage differentiation and repair cartilage defects.…”

“…Studies have shown that the intra-articular injection of verteporfin, a selective inhibitor of YAP1, can significantly maintain cartilage homeostasis in a mouse OA model and that a sustained-release system consisting of chitosan microspheres and verteporfin has significant anti-OA effects ( Zhang et al, 2020 ). Another YAP1 activity inhibitor, dasatinib, can also perform a similar function ( Nie et al, 2019 ). Dasatinib can significantly inhibit YAP1 expression, promote cartilage differentiation and repair cartilage defects.…”